Methods for iPS cell generation for basic research and clinical applications

Abstract

The induction of pluripotency can be achieved by forced expression of defined factors in somatic cells. The established cells, termed induced pluripotent stem (iPS) cells, have pluripotency and an infinite capacity for self-renewal in common with embryonic stem (ES) cells. Patient-specific iPS cells could be a useful source for drug discovery and cell transplantation therapies; however, the original method for iPS cell generation had several issues that were obstacles to their clinical application. Recent studies have brought about various improvements for iPS cell generation and uncovered several characteristics of iPS cells. Here we summarize the current status of iPS cell studies, with a focus on the improved methods that can be used to generate iPS cells, and also refer to the future challenges.

Efficient Generation of Human iPSCs by a Synthetic Self-Replicative RNA

“…However, the insertional mutagenic potential of retroviruses combined with the potential for latent reprogramming factor gene activation, especially c-MYC , all but eliminates integrative DNA-based approaches for use in regenerative medicine therapies (Okita and Yamanaka, 2011; González et al, 2011; Hussein et al, 2011; Ben-David and Benvenisty, 2011). Several methods based on DNA, RNA, miRNAs and proteins have been developed to generate integration free iPS cells, and the advantages and disadvantages have been discussed elsewhere (González et al, 2011; Hussein et al, 2011; Mochiduki and Okita, 2012; Okita and Yamanaka, 2011;). Of all these methods, RNA-based iPS cell approaches using Sendai virus (Fusaki et al, 2009), miRNAs and mRNA transfection (Warren et al, 2010) avoid potential integration problems associated with DNA-based approaches and at this point in time, appear inherently safer methods for future clinical applications.…”

Modelling Human Disease with Pluripotent Stem Cells

“…iPSC technology has moved at great speed with respect to both the refinement of derivation techniques, and the diversity of tissue types that can be reprogrammed [11]. In addition we have seen the translation of this technology to other arenas including the derivation of iPSCs from endangered species and farm animals [32].…”

“…This progress was recently acknowledged with John Gurdon and Shinya Yamanaka being awarded the Nobel Prize for their contributions to the field of reprogramming. Since the inception of induced pluripotent stem cells, the floodgates have opened with respect to refinement of the technology and its application in the generation of patient-specific models of disease [11–13]. For the first time it is now possible to generate a virtually limitless supply of highly characterised, patient-specific cells representing disease-relevant tissues [12].…”

LITERATURE REVIEW ON STEM CELL TREATMENT & ORAL SUBMUCOUS FIBROSIS (OSMF)

“…iPSCs may prove to be useful tools for drug development and modeling of diseases and in transplantation medicine. (5,6) The stem cells are also divided into totipotent stem cells (e.g., fertilized egg cell or zygote), that can generate all cell and tissue types present in an organism; Pluripotent stem cells (like embryonic stem cells) can generate the majority of cell and tissue types present in an organism; multipotent stem cells (like mesenchymal stem cells [MSC]) can generate a limited number of cell and tissue types, usually dependent on their germ layer of origin. (6) B: Properties: The main properties of stem cell, which make it different from any other specialized cells in the body are: Self-renewal, that is the ability to go through numerous cycles of cell division while maintaining their undifferentiated state; differentiation, that is the ability to differentiate into a specialized cell type and the ability to grow in vitro, in a laboratory, under a given environment.…”